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New Generation Reproductive Technologies (stem Cell-nanotechnology)

I. Advances in Embryonic and Placentalsome species CL of pregnancy is necessary
Developmenttill delivery, in others it is needed only
till placenta develops. Its complete genesis
1.Abstract:and biostimulation and bioregulatory roles
are  obscure  and  needs  further  research.
Early embryonic development needs detailed
investigations if we have to reduce embryonic10.  Dominant  follicle  dynamics
mortality. Embryonic mortality details have
been reviewed (wani, 2005). TheThe dominant follicle dynamics may be in
reproductive abnormalities have beenfocus for ovulation control. Dominant
discussed and a sequential photographicfollicle or would be bride at each ovulation
presentation of embryonic mortality haveappears around day 7 of cycle in women. It
recently been published (Khatoon et aloccurs much earlier on day- 1 to 0 in
2007). The critical period when maximumruminants (cow, sheep,goats). This time point
embryonic losses go unnoticed is the periodmay be the focus to try future selective
before attachment. Losses before actual unioninterferences in ovulation control.
of foeto-maternal tissues can be missed andAntagonist use during this phase has shown
calculations of embryonic mortality estimatesdecreasing levels of estrogens in serum
remain unaccounted for. Some recent studiesthereby indirectly hinting at dominant
of (Wani 2006, Khatoon et al 2006) havefollicle alteration, as it is one of chief
paved a new innovative model forsource of this hormone during this phase
investigation on foeto-maternal development(Fraser, 1987). Dominant follicle in its
and its attachment. This paper describes thesojourn to freedom and release at ovulation
detailed embryonic development, blastocystseems to attain some specific qualities.
elongation, formation of cotyledons withinWhy? It alone releases itself from bondages
elongated blastocyst, their ramifications andin ovary (ovulation) and rest of its
networking of vasculature and formation ofcompetitors are hindered through the process
foetal placental unit. Conversely, a detailedas atretic follicles. What make this dominant
photographic evidence of uterine receptivity,follicle to attain these qualities among a
formation of a multicoated immunologicalvast pool of follicles needs further study.
barrier- the beginning of foeto- maternal-In coming 50 years one shall possibly
barrier mechanism along with caruncleunderstand and interlock a pre-determined
formation, uterine milk formation and finallymessenger system triggering dominance in one
union of cotyledous with caruncles has beenor few follicles destined to become the new
photographically  documented.offspring of that species. This system when
decoded shall offer new opportunities in
2.Introduction:controlling ovulation both for contraception
(women) or augmentation (animals). New
Second and 3rd generation animalgenepool of dominant follicles from dead high
biotechnologies as reviewed (Wani, 2005)yielding animal ovaries would help to
are multiple ovulation, embryo-transfer,propagate  and  multiply  the  elite.
preservation, cloning, micromanipulation
splitting of embryos and nuclear transfer.Over stimulatory effect during superovulation
The 4th Generation reproductive technologiescould be avoided by use of antagonists, as
have advance beyond our imagination and nowdominant follicles seem to evade suppression,
we do have transgenic goats giving milk richthus follicular selection will prevent
in insulin. (Wani 2007). The stem cellexcessive stimulation and cystic follicles.
technology, embryonic cell culture,Follicular fluid constituents containing
pronuclear micro-injection along withoccyte awaiting for biochemical signals to
sperm-mediated, transgenesis have been in useresume (meiosis ) growth and development to
and are precisely reviewed and discussedensure timely and healthy ovulation need more
(Wani  2005,  2007).exploration (Lenton et al., 1988). The
specific signals triggering occyte release
The application of this advancement formight help in in vitro occyte maturation
diagnostic, therapeutic and economic usessuccess  rate.
especially its transfer to human medicine or
for field application in animal industry11. Development of conceptus-in vitro foetus
needs  further  research  and  evaluation.
The lean meat production needs more studies
The another fact which can be of importanceon development of conceptus. The prenatal
is the number of cells per embryos. Duringdevelopment undergoes in three stages. The
first day these may be two cells but after 8early pregnancy signals, skeletal & nervous
days the cell number increases to 120 ortissue growth, on priority. During
more. It takes 32 hours for the oocyst aftermid-gestation the hind and fore-limbs
fertilization to reach the two cell embryo.develop. During late gestation muscular
Another 13 hours are needed to develop itdevelopment along with peripheral extremities
into a four cell embryo. In another 15 hoursincluding hair develop. In order to increase
8 cell stages is reached. It is from herelean meat production, research in needed to
onwards that it takes 30 hours for a embryoinvestigate relations between hard and soft
to be converted into a Morula. This has beentissues. Maternal and foetal interactions
one of the most critical periods of embryonicneeds a probe. Some basic studies (Mufti,
development in ovine. A genematic diagram of1996) indicated prospects of manipulating
various stages of follical maturation oflean meat production (Wani, 2001, 2002 a) by
oocytes development and further developmentbio-stimulators during late gestation, which
has been reported (Wani 2005a) figuremay help, the production of muscular lambs
from web site is also show that the proteinkids at birth, besides, increasing birth
content after fertilization of 16 day oldweight. More details have been described
embryo increased by 7500 times.(Wani, 2001, 2002a) Fig.1-5). More studies in
Morphologically the embryo size varies fromfuture are needed to reveal the exact
spherical ovid to elongate after day 13thmechanism involved in critical transition of
past coitus. New protein synthesis per unitpower control from maternal to embryonic
of embryo is maximum during 8-13th day oldgenome. Application of recent techniques of
embryos. Similarly phosphorylation per unitreverse transcription, PCR and others may
of embryo is also ten times more in 8 day oldreveal  the  details  (Schultz et al., 1990).
embryos than the later stages. Glucose and
lactose production increased up to day 16th12. Slaughter  house  follicle  pool
and the maximum metabolic weight is up to
14th day past coitus. For increasedA good number of our endangered species die
productive synthesis, aminoacid requirementswhere climatic conditions are hard and harsh.
are made, through production of allanic andUnder temperate climatic conditions dead
glutonic acids by the embryos. The matobolicbodies are entrapped in snow for long. It
products get rid of amines. The signalgives a hope to revive their follicular pool.
transduction from maternal to foetal controlsOur studies on resumption of life using
have  been  discussed (Moris  et al 2001).follicles from slaughtered sheep and
fertilizing them with epididymal semen was an
3.  Dominant  follicle  dynamicspioneering attempt in this direction (Wani,
1996) (Fig 6-8). The protein content of
The new animal , products, medico-Veterinaryunfertilized ovum is around 100 mg in rabbit,
packages, profitable genetic (trait) improved27 mg in mouse (Brinister, 1967, 1971). The
DNA Chips, more resistance giving gene makesmall genetic material needs extensive
ups shall have a new role in future. For theanalysis to unveil vast informations in its
next coming super advanced technology drivencodans and anticodans. Further ovum
century, we must be prepared to use the 4thmaturation or resumption of meiosis is
generation technologies for animal Welfareinitiated during birth or shortly before
and Protective mechanisms. We thought andbirth and thus the pool of follicles remains
used Laparoscopy and ultrasonography 30 yearfixed at birth. Therefore, foetal ovary has
ago (Wani et al, 1979, Wani 1981,1982),to be the subject of further manipulations
which has now become a "buzz" word andespecially in view of its immunological
further sophisticated and modernized by myinsensitivity  (Guraya,  1985).
students across the world (Aggarwal, 2006,
Wani, 2006). Our concept of DNA finger13. Gamete  biosafety
printing of cashmere goats in 1990's
(Khatlani et al, 1995) has now crossed theBacterial counts in semen in unprecedented
barriers of technology advancement. Today,numbers effect semen fertility (Kher, and
in these pages, we shall be presenting ourDholokia, 1985), The contaminational hazards
own  results  onhave increased due to environmental
pollution, water and air contamination. Above
1. Embryonic development, formation ofall the development of resistance of known
cotyledons, blactocyst developmentbateria and other micro-organism to
ramification of vascular system, net workingantibiotics (Gupta and Maurya, 1993). We have
of blood vessels to form initial placenta,examined the semen of about 40 bulls
their union and attachment processes. Wemaintained at the Frozen semen bank,
shall describe morphological physiological,Rambirbag, Kashmir for bacterial loads in
endocrinology, histochemical and receptorsemen and tested antibiotic sensitivity
analysis to support our observations. This is(Koul,  2002).
5th generation reproductive technology base
for  development  of  in-vitro-foeto  models.Presence of bacteria in large numbers in
semen, effects its quality and fertility
For 6th - 10th generation reproductive(Kher and Dholokia, 1985). The source of
scientists we shall present theoreticalcontamination of semen vary from inflammatory
hypothesis  a  vision  -2050  in the form of.foci and lesions on penile apparatus,
contaminations on equipments, appliances,
2. Use of stem-cell, germ-cell and Nano-diluters, extenders, air and laboratory
technologies for production of disease free,environment. These contamination hazards have
highly prolific reproductive gametes, theirincreased with environmental pollution. Many
preservation and conservation. The use ofbacteria having developed resistance to
these and other molecular and new-generationroutine antibiotics used (Gupta and Maurya
DNA-related bio-diagnostics shall also be1993) in semen extenders has made
discussed.inseminations more critical and vulnerable.
More studies in this direction are needed to
4.  Uterine  morphological  changes:develop a bio-safe and infection free
germplasm pool of frozen and fresh semen in
The early pregnancy uterine morphologicalfuture.
land marks were described only externally per
rectum in bovine (Zamznis, 1976) some of14. Foeto-maternal  interaction
its significant features have been summarized
in a booklet (Wani, 1986). It describesThe role of endometrial stromal cells during
easy palpable signs of early pregnancy inpre-implantation stage needs further
bovine and compares its observations withresearch. Some studies on ultrastructure of
equines, sheep and goats. Some of theendometrium (Brinsfield et al., 1974) are
earliest indications of pregnancy by physicalstill not sufficient to answer the questions.
examination were:i) Presence of corpus luteumWhat is the role of immune cells and
of pregnancy early 15 day post-coitusii)endometrial cells in building of the
Asymmetry of uterine horns 30-35 days postimmunological barrier to protect the foetus.
coitusiii) Fluctuation around 40-49 days inFoetal biology is abarren field of research
cattle. Thereafter the advanced pregnancyin the world. After few initial steps by Dr.
signs like foetal membrance feel and slips.Moris Young in UK and D. Srivastava in
Palpation of placentomes and full foetal areIndia, only scattered information is
known  foetal  membrane  .available on the subject (Wani and Buchoo,
1990; Mufti, 1996) . Much is still to be
Various developments of foetal signs haveunveiled by 2050. The placental hormone
been described (Wani, 1986) . The monthinfluence mammary development and milk
wise morphological assessment of foetus inproduction of the dam. Foetal membranes serve
sheep has been reported (Mufti, 1995, Muftias Chief source of nutritive exchanges. The
et al, 1997) . Various reviews reports andarrangement of cotyledons, caruncles,
books now describe events in development ofpositioning of single and twin conceptus,
foetus  (Wani,  2005,  2006,  2007).indicate complex mechanisms with common
roots. (Mufti et al., 2000) These
5. Development of conceptus-in vitromechanisms of foeto-maternal interactions and
foetusrelationships emphasize proper foetal number
diagnosis and corresponding
Pregnancy is an intimate association betweenmaterno-nutritional  management  strategies.
mother and foetus. The protective covering on
maternal caruncles and over wrap of foetal15.  Nanotechnology:
cotyledons with Blastocyst membranes,
transformed into foetal membranes give theIn the proceeding pages we have seen the role
"Homograph" status to foetus. The mechanismof biotechnology in animal reproduction. The
of ruminant uteri has been discussed (Gray etcoming generations will harvest the gains in
2002). Its more recent findings andthe form of more micron-level-accurate
attachments are the topic of one of ourtechnology being replaced with
students.nanogram-level-invisible technology- the
nanotechnology. Now even converging
We have tried to univel many obscuretechnologies are shaped resulting from merger
mechanism on blastocyst formation,of Nano- science and Nanotechnology with
plancentation and development of cotyledons.biomedicine, bio-sciences, biotechnology,
The protective mucin coverings over uterinegenetic engineering and information
caruncles and their union with cotyledons hastechnology.
been  snaped.
The cognitive sciences like Neuro- sciences,
The internal changes have been correlatedcomputing, communication combined with
with biochemical, morphological andnanotechnology may be so mystic and
histochemical and histoanatomic feature ofdeterministic that science shows will appear
the  gravid  and  non-gravid  uteri.as "Majic shows" . This new technology
wave" may define our spiritualistic
6. Foeto-maternal  interaction:wavelengths. Many unsolved or resolved facts
on birth and genetic make up shall send such
The estrogen, progesterone in foetal andmessages as was known of Sagic saints, peers
maternal blood have been estimated along withand  Fakirs.
estrogen progesterone receptor concentrations
in  the  pregnancy  and  non pregnant states.The birth of a technology needs patience of
public and policy makers. In the past
Endometrial receptivity has been discussed"Michael Farady" - the inventor of electric
(Huct-Hudson, et al 1989, Wani 1996, Wani,generator, was asked by the then Chancellor
2003, 205, 2006). Proliferation of uterineof Exchequer, as to what use is the
cells is influenced by E2 and P4. Theseelectricity you are generating. He argues but
hormones have their receptors in uteri calledno argument convinced the accountant.
as E2 (ER) , P4 (PR) receptors (Lee and DemyoFinally, he promised him that the day will
2004) we have also investigated the same andcome when you can tax it. Today a remote
are being analysed. Our studies indicatedestination on this earth demands 24 hr
certain changes which project a new mechanismelectricity. They never argue its price, but
of foeto-maternal unions (Khatoon and Wani,its availability as savoir to life be it
2007).medical, veterinary or any other field. Thus
like Farady I profess and advocate
Various growth factors described (Wani 2006)experiments on use of nanotechnology in
may be helpful in future for invitro-foetalanimal reproduction today. It may sound
growth.strange, but yes in my life I shall fore see
a role of nanotechnology in Animal
7. Reproductive  managementreproduction.
The reproductive management strategies to16.  What  is  Nanotechnology:
enhance meat production could safely be
divided into two kinds:i. TraditionalTechnology related to features of nanometer
management: Animal are still managed underscale. Thus we will unveil gametes in minute
zero- input/ migratory /tribal or extensivedetails; sperm head colliding zona-pellucida
management system. The application of aboveshall be filmed. The radial blast of cortical
described methods will be limited. They mayreaction shall be known. All chemical
indirectly help in this system by better siremessengers  shall  be unveiled at nano-level.
availability.ii. Modern management:
Zero-grazing, stall-fed or semi-intensive,The shape of gamete, Zygote, embryo and
commercial goat farming system may use thefoetal development at nanolevel shall bring
above methods more and more to produce highforth new additions to our knowledge. The use
quality germplasm. They may use variety ofof such information through use of computer
improved reproductive technique or reportedlanguages, both hard and soft shall make this
(Wani,1996; Wani et al., 200a; Wani et al.,knowledge dispensable and useable at
1998; Mufti et al., 1997; Wani and Sahani,remote-corners of the Universe. Combing
1981; Wani et al., 1986, 1987, 1988; Wani andcognitive, mythical and mysterious happening
Buchoo, 1991; Buchoo and Wani, 1991; Wani,within womb could be registered and
1989, Wani, 1995; Wani, 1984a and b; Wani andtranslated  via  nano-ultra-sonography.
Sahani,  1980a).
Thus futuristic view of nanotechnology will
8. Enhancement  of  fertilitybe nanographic follow of information and has
been  described  (  Mordini,  E,  2007).
Desired fertility objectives have been
discussed (Wani, 2001). IncreasedNanotechnology may allow us to send tiny nano
prolificacy, early rebreeding, safe/cleanmicroprocessors into the animal womb to send
parturition, set-time animal crop, elite geneus signals which we wish to know for
introduction are some of the desiredprevention of diseases. The
fertility enhancement goals. The constraintsstem-cell-germ-cell technology combine with
in achieving desired objectives in smallNanotechnology shall bring new hope and
ruminant livestock sector are the problems offuture for the propapagation of
inbreeding, reproductive losses (Wani, etGene-technology-ETT- A-I Combine. It may
al., 1980; 1981; 1994), pre-pubertal mating,sound like sending a wave of spirit to
indiscriminate or illicit mating, venerealfertilize an "egg" - thus proving the "God
diseases, nutritional and other stresses.Word in Quran of creating the Chirst
These above referred factors reduce fertilitythrough mystic spirit. This and other
status of our ruminants especially underconvergence of known reproductive
transhumane  migratory  systems.technologies  are  challenge  to us in future
These constraints mentioned could overcome byNanaoparticels developed can be agents which
adopting enhancement strategies (Wani, 2001),would use blood and carry durgs for curing
which include exchange of sires by tribal,disease at molecular levels. Thus a new
Gujjar, Bakarwals and Chopans to reducemechanism of targeting infective agents of
inbreeding. This could be ensured throughsmaller  dimension  could  be  a possibility.
legislature or forced castrations and
introduction of sires from outside the flock.17.  Stem-cell-Technology:
The pregnancy detection methods described
(Wani, 1981) and later on used extensive inStem- cells are undifferentiated cells. The
J&K under farm condition (Wani et al., 1998;embryo at morula stages have undifferentiated
Wani, 1989) needs adoption as a routine inblastomeres. These cells can transform
each flock for better care of offspring andthemselves into 200 or more cell type, which
dam.could be used to repair or regenerate new
desired cells. This advanced cell research
9. Corpora  luteal  biosensitivity.may help diseased people like those suffering
from cancer, Alzheimer's disease syndrome,
Corpus luteum growth and development is theParkinsons  disease  and  even  paralysis.
first earliest pregnancy signal along with
maternal recognition factors discussed (Wani,The stem cells are derived from 4-5 day old
1996). The corpus luteum development andembryos or fertilized cells. The
receptivity to PGF2 alpha is now known. Thestem-cell-technology has a new role to play
ascending and descending corpora lutea arein  Animal  reproduction.
insensitive to PGF2 alpha. What makes them
insensitive and what resurges theirFirstly, the stem-cell source could be pooled
sensitivity need exploration. May be inout of slaughter-house oocytes or from the
future, CL insensitivity might have directvast pool of embryos hatching out in many
effects on survival of the conceptus. Doesanimal  species.
conceptus make CL more sensitive to PGF2
alpha? Or in other words, does the gravid CLSecondly, the cell source could be obtained
and non-gravid CL behave similarly orfrom foetal culture and if
differently to PGF2 alpha. Many suchnew-grown-immunoprotectant cell of foetus
questions vis-Ã -vis the ovarian morphology,could help or provide germ-cell specific to
uterine environment and behavioral patternsany particular organ, the repair of organs
need to be studied and co-related to developcould be a new revolution.
a manipulative and curative safe system. In



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