| I. Advances in Embryonic and Placental Development | | | | environment and behavioral patterns need to be |
| 1.Abstract: | | | | studied and co-related to develop a manipulative and |
| Early embryonic development needs detailed | | | | curative safe system. In some species CL of |
| investigations if we have to reduce embryonic | | | | pregnancy is necessary till delivery, in others it is |
| mortality. Embryonic mortality details have been | | | | needed only till placenta develops. Its complete |
| reviewed (wani, 2005). The reproductive abnormalities | | | | genesis and biostimulation and bioregulatory roles are |
| have been discussed and a sequential photographic | | | | obscure and needs further research. |
| presentation of embryonic mortality have recently | | | | 10. Dominant follicle dynamics |
| been published (Khatoon et al 2007). The critical | | | | The dominant follicle dynamics may be in focus for |
| period when maximum embryonic losses go unnoticed | | | | ovulation control. Dominant follicle or would be bride |
| is the period before attachment. Losses before actual | | | | at each ovulation appears around day 7 of cycle in |
| union of foeto-maternal tissues can be missed and | | | | women. It occurs much earlier on day- 1 to 0 in |
| calculations of embryonic mortality estimates remain | | | | ruminants (cow, sheep,goats). This time point may be |
| unaccounted for. Some recent studies of (Wani 2006, | | | | the focus to try future selective interferences in |
| Khatoon et al 2006) have paved a new innovative | | | | ovulation control. Antagonist use during this phase has |
| model for investigation on foeto-maternal | | | | shown decreasing levels of estrogens in serum |
| development and its attachment. This paper | | | | thereby indirectly hinting at dominant follicle alteration, |
| describes the detailed embryonic development, | | | | as it is one of chief source of this hormone during |
| blastocyst elongation, formation of cotyledons within | | | | this phase (Fraser, 1987). Dominant follicle in its |
| elongated blastocyst, their ramifications and | | | | sojourn to freedom and release at ovulation seems |
| networking of vasculature and formation of foetal | | | | to attain some specific qualities. Why? It alone |
| placental unit. Conversely, a detailed photographic | | | | releases itself from bondages in ovary (ovulation) and |
| evidence of uterine receptivity, formation of a | | | | rest of its competitors are hindered through the |
| multicoated immunological barrier- the beginning of | | | | process as atretic follicles. What make this dominant |
| foeto- maternal- barrier mechanism along with | | | | follicle to attain these qualities among a vast pool of |
| caruncle formation, uterine milk formation and finally | | | | follicles needs further study. In coming 50 years one |
| union of cotyledous with caruncles has been | | | | shall possibly understand and interlock a |
| photographically documented. | | | | pre-determined messenger system triggering |
| 2.Introduction: | | | | dominance in one or few follicles destined to become |
| Second and 3rd generation animal biotechnologies as | | | | the new offspring of that species. This system when |
| reviewed (Wani, 2005) are multiple ovulation, | | | | decoded shall offer new opportunities in controlling |
| embryo-transfer, preservation, cloning, | | | | ovulation both for contraception (women) or |
| micromanipulation splitting of embryos and nuclear | | | | augmentation (animals). New genepool of dominant |
| transfer. The 4th Generation reproductive | | | | follicles from dead high yielding animal ovaries would |
| technologies have advance beyond our imagination | | | | help to propagate and multiply the elite. |
| and now we do have transgenic goats giving milk rich | | | | Over stimulatory effect during superovulation could |
| in insulin. (Wani 2007). The stem cell technology, | | | | be avoided by use of antagonists, as dominant |
| embryonic cell culture, pronuclear micro-injection along | | | | follicles seem to evade suppression, thus follicular |
| with sperm-mediated, transgenesis have been in use | | | | selection will prevent excessive stimulation and cystic |
| and are precisely reviewed and discussed (Wani | | | | follicles. Follicular fluid constituents containing occyte |
| 2005, 2007). | | | | awaiting for biochemical signals to resume (meiosis ) |
| The application of this advancement for diagnostic, | | | | growth and development to ensure timely and |
| therapeutic and economic uses especially its transfer | | | | healthy ovulation need more exploration (Lenton et |
| to human medicine or for field application in animal | | | | al., 1988). The specific signals triggering occyte release |
| industry needs further research and evaluation. | | | | might help in in vitro occyte maturation success rate. |
| The another fact which can be of importance is the | | | | 11. Development of conceptus-in vitro foetus |
| number of cells per embryos. During first day these | | | | The lean meat production needs more studies on |
| may be two cells but after 8 days the cell number | | | | development of conceptus. The prenatal |
| increases to 120 or more. It takes 32 hours for the | | | | development undergoes in three stages. The early |
| oocyst after fertilization to reach the two cell | | | | pregnancy signals, skeletal & nervous tissue growth, |
| embryo. Another 13 hours are needed to develop it | | | | on priority. During mid-gestation the hind and |
| into a four cell embryo. In another 15 hours 8 cell | | | | fore-limbs develop. During late gestation muscular |
| stages is reached. It is from here onwards that it | | | | development along with peripheral extremities |
| takes 30 hours for a embryo to be converted into a | | | | including hair develop. In order to increase lean meat |
| Morula. This has been one of the most critical periods | | | | production, research in needed to investigate relations |
| of embryonic development in ovine. A genematic | | | | between hard and soft tissues. Maternal and foetal |
| diagram of various stages of follical maturation of | | | | interactions needs a probe. Some basic studies (Mufti, |
| oocytes development and further development has | | | | 1996) indicated prospects of manipulating lean meat |
| been reported (Wani 2005a) figure from web site is | | | | production (Wani, 2001, 2002 a) by bio-stimulators |
| also show that the protein content after fertilization | | | | during late gestation, which may help, the production |
| of 16 day old embryo increased by 7500 times. | | | | of muscular lambs/kids at birth, besides, increasing |
| Morphologically the embryo size varies from spherical | | | | birth weight. More details have been described (Wani, |
| ovid to elongate after day 13th past coitus. New | | | | 2001, 2002a) Fig.1-5). More studies in future are |
| protein synthesis per unit of embryo is maximum | | | | needed to reveal the exact mechanism involved in |
| during 8-13th day old embryos. Similarly | | | | critical transition of power control from maternal to |
| phosphorylation per unit of embryo is also ten times | | | | embryonic genome. Application of recent techniques |
| more in 8 day old embryos than the later stages. | | | | of reverse transcription, PCR and others may reveal |
| Glucose and lactose production increased up to day | | | | the details (Schultz et al., 1990). |
| 16th and the maximum metabolic weight is up to | | | | 12. Slaughter house follicle pool |
| 14th day past coitus. For increased productive | | | | A good number of our endangered species die where |
| synthesis, aminoacid requirements are made, through | | | | climatic conditions are hard and harsh. Under |
| production of allanic and glutonic acids by the | | | | temperate climatic conditions dead bodies are |
| embryos. The matobolic products get rid of amines. | | | | entrapped in snow for long. It gives a hope to revive |
| The signal transduction from maternal to foetal | | | | their follicular pool. Our studies on resumption of life |
| controls have been discussed (Moris et al 2001). | | | | using follicles from slaughtered sheep and fertilizing |
| 3. Dominant follicle dynamics | | | | them with epididymal semen was an pioneering |
| The new animal , products, medico-Veterinary | | | | attempt in this direction (Wani, 1996) (Fig 6-8). The |
| packages, profitable genetic (trait) improved DNA | | | | protein content of unfertilized ovum is around 100 mg |
| Chips, more resistance giving gene make ups shall | | | | in rabbit, 27 mg in mouse (Brinister, 1967, 1971). The |
| have a new role in future. For the next coming super | | | | small genetic material needs extensive analysis to |
| advanced technology driven century, we must be | | | | unveil vast informations in its codans and anticodans. |
| prepared to use the 4th generation technologies for | | | | Further ovum maturation or resumption of meiosis is |
| animal Welfare and Protective mechanisms. We | | | | initiated during birth or shortly before birth and thus |
| thought and used Laparoscopy and ultrasonography | | | | the pool of follicles remains fixed at birth. Therefore, |
| 30 year ago (Wani et al, 1979, Wani 1981,1982), | | | | foetal ovary has to be the subject of further |
| which has now become a "buzz" word and further | | | | manipulations especially in view of its immunological |
| sophisticated and modernized by my students across | | | | insensitivity (Guraya, 1985). |
| the world (Aggarwal, 2006, Wani, 2006). Our concept | | | | 13. Gamete biosafety |
| of DNA finger printing of cashmere goats in 1990's | | | | Bacterial counts in semen in unprecedented numbers |
| (Khatlani et al, 1995) has now crossed the barriers of | | | | effect semen fertility (Kher, and Dholokia, 1985), The |
| technology advancement. Today, in these pages, we | | | | contaminational hazards have increased due to |
| shall be presenting our own results on | | | | environmental pollution, water and air contamination. |
| 1. Embryonic development, formation of cotyledons, | | | | Above all the development of resistance of known |
| blactocyst development ramification of vascular | | | | bateria and other micro-organism to antibiotics (Gupta |
| system, net working of blood vessels to form initial | | | | and Maurya, 1993). We have examined the semen of |
| placenta, their union and attachment processes. We | | | | about 40 bulls maintained at the Frozen semen bank, |
| shall describe morphological physiological, | | | | Rambirbag, Kashmir for bacterial loads in semen and |
| endocrinology, histochemical and receptor analysis to | | | | tested antibiotic sensitivity (Koul, 2002). |
| support our observations. This is 5th generation | | | | Presence of bacteria in large numbers in semen, |
| reproductive technology base for development of | | | | effects its quality and fertility (Kher and Dholokia, |
| in-vitro-foeto models. | | | | 1985). The source of contamination of semen vary |
| For 6th - 10th generation reproductive scientists we | | | | from inflammatory foci and lesions on penile |
| shall present theoretical hypothesis a vision -2050 in | | | | apparatus, contaminations on equipments, appliances, |
| the form of. | | | | diluters, extenders, air and laboratory environment. |
| 2. Use of stem-cell, germ-cell and Nano- technologies | | | | These contamination hazards have increased with |
| for production of disease free, highly prolific | | | | environmental pollution. Many bacteria having |
| reproductive gametes, their preservation and | | | | developed resistance to routine antibiotics used |
| conservation. The use of these and other molecular | | | | (Gupta and Maurya 1993) in semen extenders has |
| and new-generation DNA-related bio-diagnostics shall | | | | made inseminations more critical and vulnerable. More |
| also be discussed. | | | | studies in this direction are needed to develop a |
| 4. Uterine morphological changes: | | | | bio-safe and infection free germplasm pool of frozen |
| The early pregnancy uterine morphological land marks | | | | and fresh semen in future. |
| were described only externally per rectum in bovine | | | | 14. Foeto-maternal interaction |
| (Zamznis, 1976) some of its significant features have | | | | The role of endometrial stromal cells during |
| been summarized in a booklet (Wani, 1986). It | | | | pre-implantation stage needs further research. Some |
| describes easy palpable signs of early pregnancy in | | | | studies on ultrastructure of endometrium (Brinsfield et |
| bovine and compares its observations with equines, | | | | al., 1974) are still not sufficient to answer the |
| sheep and goats. Some of the earliest indications of | | | | questions. What is the role of immune cells and |
| pregnancy by physical examination were:i) Presence | | | | endometrial cells in building of the immunological barrier |
| of corpus luteum of pregnancy early 15 day | | | | to protect the foetus. Foetal biology is abarren field |
| post-coitusii) Asymmetry of uterine horns 30-35 days | | | | of research in the world. After few initial steps by Dr. |
| post coitusiii) Fluctuation around 40-49 days in cattle. | | | | Moris Young in UK and D. Srivastava in India, only |
| Thereafter the advanced pregnancy signs like foetal | | | | scattered information is available on the subject |
| membrance feel and slips. Palpation of placentomes | | | | (Wani and Buchoo, 1990; Mufti, 1996) . Much is still to |
| and full foetal are known foetal membrane . | | | | be unveiled by 2050. The placental hormone influence |
| Various developments of foetal signs have been | | | | mammary development and milk production of the |
| described (Wani, 1986) . The month wise | | | | dam. Foetal membranes serve as Chief source of |
| morphological assessment of foetus in sheep has | | | | nutritive exchanges. The arrangement of cotyledons, |
| been reported (Mufti, 1995, Mufti et al, 1997) . | | | | caruncles, positioning of single and twin conceptus, |
| Various reviews reports and books now describe | | | | indicate complex mechanisms with common roots. |
| events in development of foetus (Wani, 2005, 2006, | | | | (Mufti et al., 2000) These mechanisms of |
| 2007). | | | | foeto-maternal interactions and relationships |
| 5. Development of conceptus-in vitro foetus | | | | emphasize proper foetal number diagnosis and |
| Pregnancy is an intimate association between mother | | | | corresponding materno-nutritional management |
| and foetus. The protective covering on maternal | | | | strategies. |
| caruncles and over wrap of foetal cotyledons with | | | | 15. Nanotechnology: |
| Blastocyst membranes, transformed into foetal | | | | In the proceeding pages we have seen the role of |
| membranes give the "Homograph" status to foetus. | | | | biotechnology in animal reproduction. The coming |
| The mechanism of ruminant uteri has been discussed | | | | generations will harvest the gains in the form of |
| (Gray et 2002). Its more recent findings and | | | | more micron-level-accurate technology being replaced |
| attachments are the topic of one of our students. | | | | with nanogram-level-invisible technology- the |
| We have tried to univel many obscure mechanism on | | | | nanotechnology. Now even converging technologies |
| blastocyst formation, plancentation and development | | | | are shaped resulting from merger of Nano- science |
| of cotyledons. The protective mucin coverings over | | | | and Nanotechnology with biomedicine, bio-sciences, |
| uterine caruncles and their union with cotyledons has | | | | biotechnology, genetic engineering and information |
| been snaped. | | | | technology. |
| The internal changes have been correlated with | | | | The cognitive sciences like Neuro- sciences, |
| biochemical, morphological and histochemical and | | | | computing, communication combined with |
| histoanatomic feature of the gravid and non-gravid | | | | nanotechnology may be so mystic and deterministic |
| uteri. | | | | that science shows will appear as "Majic shows" . This |
| 6. Foeto-maternal interaction: | | | | new technology wave" may define our spiritualistic |
| The estrogen, progesterone in foetal and maternal | | | | wavelengths. Many unsolved or resolved facts on |
| blood have been estimated along with estrogen | | | | birth and genetic make up shall send such messages |
| progesterone receptor concentrations in the | | | | as was known of Sagic saints, peers and Fakirs. |
| pregnancy and non pregnant states. | | | | The birth of a technology needs patience of public |
| Endometrial receptivity has been discussed | | | | and policy makers. In the past "Michael Farady" - the |
| (Huct-Hudson, et al 1989, Wani 1996, Wani, 2003, | | | | inventor of electric generator, was asked by the |
| 205, 2006). Proliferation of uterine cells is influenced | | | | then Chancellor of Exchequer, as to what use is the |
| by E2 and P4. These hormones have their receptors | | | | electricity you are generating. He argues but no |
| in uteri called as E2 (ER) , P4 (PR) receptors (Lee and | | | | argument convinced the accountant. Finally, he |
| Demyo 2004) we have also investigated the same | | | | promised him that the day will come when you can |
| and are being analysed. Our studies indicate certain | | | | tax it. Today a remote destination on this earth |
| changes which project a new mechanism of | | | | demands 24 hr electricity. They never argue its price, |
| foeto-maternal unions (Khatoon and Wani, 2007). | | | | but its availability as savoir to life be it medical, |
| Various growth factors described (Wani 2006) may | | | | veterinary or any other field. Thus like Farady I |
| be helpful in future for invitro-foetal growth. | | | | profess and advocate experiments on use of |
| 7. Reproductive management | | | | nanotechnology in animal reproduction today. It may |
| The reproductive management strategies to enhance | | | | sound strange, but yes in my life I shall fore see a |
| meat production could safely be divided into two | | | | role of nanotechnology in Animal reproduction. |
| kinds:i. Traditional management: Animal are still | | | | 16. What is Nanotechnology: |
| managed under zero- input/ migratory /tribal or | | | | Technology related to features of nanometer scale. |
| extensive management system. The application of | | | | Thus we will unveil gametes in minute details; sperm |
| above described methods will be limited. They may | | | | head colliding zona-pellucida shall be filmed. The radial |
| indirectly help in this system by better sire availability.ii. | | | | blast of cortical reaction shall be known. All chemical |
| Modern management: Zero-grazing, stall-fed or | | | | messengers shall be unveiled at nano-level. |
| semi-intensive, commercial goat farming system may | | | | The shape of gamete, Zygote, embryo and foetal |
| use the above methods more and more to produce | | | | development at nanolevel shall bring forth new |
| high quality germplasm. They may use variety of | | | | additions to our knowledge. The use of such |
| improved reproductive technique or reported | | | | information through use of computer languages, both |
| (Wani,1996; Wani et al., 200a; Wani et al., 1998; Mufti | | | | hard and soft shall make this knowledge dispensable |
| et al., 1997; Wani and Sahani, 1981; Wani et al., 1986, | | | | and useable at remote-corners of the Universe. |
| 1987, 1988; Wani and Buchoo, 1991; Buchoo and | | | | Combing cognitive, mythical and mysterious happening |
| Wani, 1991; Wani, 1989, Wani, 1995; Wani, 1984a and | | | | within womb could be registered and translated via |
| b; Wani and Sahani, 1980a). | | | | nano-ultra-sonography. |
| 8. Enhancement of fertility | | | | Thus futuristic view of nanotechnology will be |
| Desired fertility objectives have been discussed | | | | nanographic follow of information and has been |
| (Wani, 2001). Increased prolificacy, early rebreeding, | | | | described ( Mordini, E, 2007). |
| safe/clean parturition, set-time animal crop, elite gene | | | | Nanotechnology may allow us to send tiny nano |
| introduction are some of the desired fertility | | | | microprocessors into the animal womb to send us |
| enhancement goals. The constraints in achieving | | | | signals which we wish to know for prevention of |
| desired objectives in small ruminant livestock sector | | | | diseases. The stem-cell-germ-cell technology combine |
| are the problems of inbreeding, reproductive losses | | | | with Nanotechnology shall bring new hope and future |
| (Wani, et al., 1980; 1981; 1994), pre-pubertal mating, | | | | for the propapagation of Gene-technology-ETT- A-I |
| indiscriminate or illicit mating, venereal diseases, | | | | Combine. It may sound like sending a wave of spirit |
| nutritional and other stresses. These above referred | | | | to fertilize an "egg" - thus proving the "God Word in |
| factors reduce fertility status of our ruminants | | | | Quran of creating the Chirst through mystic spirit. |
| especially under transhumane migratory systems. | | | | This and other convergence of known reproductive |
| These constraints mentioned could overcome by | | | | technologies are challenge to us in future |
| adopting enhancement strategies (Wani, 2001), which | | | | Nanaoparticels developed can be agents which would |
| include exchange of sires by tribal, Gujjar, Bakarwals | | | | use blood and carry durgs for curing disease at |
| and Chopans to reduce inbreeding. This could be | | | | molecular levels. Thus a new mechanism of targeting |
| ensured through legislature or forced castrations and | | | | infective agents of smaller dimension could be a |
| introduction of sires from outside the flock. The | | | | possibility. |
| pregnancy detection methods described (Wani, 1981) | | | | 17. Stem-cell-Technology: |
| and later on used extensive in J&K under farm | | | | Stem- cells are undifferentiated cells. The embryo at |
| condition (Wani et al., 1998; Wani, 1989) needs | | | | morula stages have undifferentiated blastomeres. |
| adoption as a routine in each flock for better care of | | | | These cells can transform themselves into 200 or |
| offspring and dam. | | | | more cell type, which could be used to repair or |
| 9. Corpora luteal biosensitivity. | | | | regenerate new desired cells. This advanced cell |
| Corpus luteum growth and development is the first | | | | research may help diseased people like those |
| earliest pregnancy signal along with maternal | | | | suffering from cancer, Alzheimer's disease syndrome, |
| recognition factors discussed (Wani, 1996). The | | | | Parkinsons disease and even paralysis. |
| corpus luteum development and receptivity to PGF2 | | | | The stem cells are derived from 4-5 day old |
| alpha is now known. The ascending and descending | | | | embryos or fertilized cells. The stem-cell-technology |
| corpora lutea are insensitive to PGF2 alpha. What | | | | has a new role to play in Animal reproduction. |
| makes them insensitive and what resurges their | | | | Firstly, the stem-cell source could be pooled out of |
| sensitivity need exploration. May be in future, CL | | | | slaughter-house oocytes or from the vast pool of |
| insensitivity might have direct effects on survival of | | | | embryos hatching out in many animal species. |
| the conceptus. Does conceptus make CL more | | | | Secondly, the cell source could be obtained from |
| sensitive to PGF2 alpha? Or in other words, does the | | | | foetal culture and if new-grown-immunoprotectant |
| gravid CL and non-gravid CL behave similarly or | | | | cell of foetus could help or provide germ-cell specific |
| differently to PGF2 alpha. Many such questions | | | | to any particular organ, the repair of organs could be |
| vis-Ã -vis the ovarian morphology, uterine | | | | a new revolution. |