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Biomedical subjects

K Sailaja

Publications and source records attributed to K Sailaja.

7 recordsLinked to original sources

Cell surface molecules (NCAM and L1) in intrastriatal transplants of embryonic mesencephalon in rats.

Cell surface molecules, NCAM and L1, reported to have a role in synaptogenesis, growth and fasciculation of the neurites in the brain, were traced in the embryonic nigral transplants in the host striatum of adult rats. Substantia nigra of five, 15 and 25 postnatal days were also examined for the same molecules. Tyrosine hydroxylase label was used as a marker to localize the nigral neurons and glial fibrillary acidic protein to detect if glial scar present. In the control as well as transplants large neurons had expressed tyrosine hydroxylase. By 15th postnatal day tyrosine hydroxylase neurons appeared mature and were scattered, suggesting a well-formed neuropil. NCAM and L1 reaction was seen as a peripheral rim in most of the cells on the fifth postnatal day. The reaction was mainly in relation to the large cells and more extensive on the 15th day. Thereafter on the 25th day, activity was negligible. Large neurons demonstrated strong reactivity for NCAM and L1 during early post-transplantation days. After 30 days only smaller cells were reactive, many of which could be identified as neurons. Strong reaction for these molecules was present only until 60 days, though faint reaction could be detected even on the 90th day. These observations indicate that the growth promoting molecules, the type seen in the neonatal period, can be detected normally only until the neurons mature. Prolonged expression of these molecules by the grafted neurons indicate delay in the maturation of these cells due to absence of adequate target sites for synaptic connections. Some of the smaller cells expressing these molecules after 30 days of transplantation could be astroglia, either proliferating or reactive.

Animals

Ultrastructure of developing substantia nigra in humans.

Electron microscopy of the maturing neurons and developing and maturing synapses in the substantia nigra of 14 human embryos/foetuses of 8-24 weeks of gestation are reported. At 8 weeks, cells were immature with very little cytoplasm and cellular organelles. Contact sites of processes appeared more electron dense than the other areas. At 12 weeks, many of the cells had acquired more cytoplasm and cellular organelles and could be identified as neurons. Asymmetric synapses with clear, round synaptic vesicles also were identifiable at this age. Such synapses, first to appear in the developing substantia nigra, are reported to be formed by recurrent collateral nigro-striatal fibres. Substance P fibres from the striatum also are contributing to this type of synapse. At 15-16 weeks, not only was the number of such synapses increased, but many appeared morphologically mature. Symmetric synapses having clear round vesicles along with a few dense core vesicles also appeared at this stage, suggesting striatal input. By 24 weeks of gestation, most of the neurons had cytological features comparable to that of the mature neurons. There was an increase in the total number of synapses and the individual variety from 15 to 24 weeks of gestation. The present study indicates that synaptogenesis starts at 8 weeks and continues beyond 24 weeks of gestation.

Cytoplasm

Long-term nigral transplants in rat striatum: an electron microscopic study.

The substantia nigra of gestation day 14 was transplanted into the striatum of 3-4-month-old rats to investigate the transplants ultrastructurally at the end of 2 years, as a follow-up to our previous studies. Transplants were of small size in all 10 specimens taken for this study. The changes observed in the transplant and in the interface region with the host striatum were: thickening of the blood vessel walls, perivascular cuffing with lymphocytes and macrophages loaded with tissue debris, degenerating neurons and hypertrophied astroglia containing dense granules indicating ageing or reaction to degeneration and glial processes. The number of surviving neurons in the transplants was small. These were smaller in size and had very few intracytoplasmic membraneous organelles. A higher content of intracytoplasmic ageing lipofuscin pigment was present than in host neurons and age-matched nigral neurons. Synapses were few, and their number varied among transplants. Generally, the synapses were at the interface with the host tissue. The changes observed in all the 2-year-old transplants suggest premature ageing or a slow rejection process. Slow rejection is a possibility, because these rats are only stock-bred, not inbred, and hence they are not completely immunologically compatible.

Animals

Developing substantia nigra in human: a qualitative study.

Midbrains from 43 fresh human embryos and fetuses at 8-22 weeks of gestation were processed for routine histology, Golgi staining, tyrosine hydroxylase (TH) immunolabelling and retrograde tracing with the fluorescent dye DiI. Cells were immature and densely packed between 8 and 10 weeks. By 13 weeks cells could be identified as neurons and glia. Neurons matured gradually and achieved adult characteristics by 20-22 weeks. Neurons in the paramedian regions of the tegmentum, raphae region and substantia nigra were positive for TH from 13 weeks onwards, the earliest age group used for this technique. The presence of TH-positive neurons in the paramedian part of the tegmentum until 18 weeks and radial glial fibers extending from the aqueductal lining to the ventral part until 20 weeks were suggestive of migration of neurons to the ventral mesencephalon region. DiI labelling of the neurons and fibers of ipsilateral nigra from the caudate as early as 10 weeks demonstrated early nigrostriatal connections. The mature nature of the neurons appeared only by 13 weeks by this method. The present study shows that the nigral neurons in the human migrate and mature until mid-gestation. The nigrostriatal connection at 10 weeks suggests a trophic influence of nigra on the proliferating and maturing neurons of the striatum.

Carbocyanines

Fetal dopaminergic neurons transplanted to the normal striatum of neonatal or adult rats and to the denervated striatum of adult rats.

Fetal ventral mesencephalon from the 15th gestational day was grafted into the striatum of neonatal and adult rats. In one group of adult rats, fetal nigra was transplanted into normal striatum. In a second group, the tissue was transplanted at sites where dopaminergic fibers were denervated with 6-hydroxydopamine. The behavior of the dopaminergic neurons and glial reactions were studied by staining with cresyl violet to localize the transplants and by immunolabeling tyrosine hydroxylase (TH) and glial fibrillary acidic protein. In normal adults, the transplants were small. At the edge of the transplants, TH-positive neurons were packed into clusters, and an interface without any significant crossover of TH-positive fibers was present. Glial reaction was minimal in and around the transplant. In the denervated striatum, transplants were generally larger than those in normal striatum and surrounded by a glial scar. TH-positive neurons were both closely packed and loosely arranged at the periphery of the transplants. Processes could be clearly defined and could be traced to the adjacent host striatum through the TH-free denervated area. In neonates, the transplants were large and at times extended beyond the striatum. Most TH-positive neurons were arranged linearly along the periphery of the transplant. Cell bodies were widely separated and a well-developed neuropil was present. Fibers from the transplant mingled freely with the host striatum without any interface. In all three transplant groups, tracing the TH-positive neurites was easy because they were thicker and coarser than other elements. No apparent glial reaction occurred in the neonates. Thus, the growth and maturation of dopaminergic neurons seemed to vary in different environments. The most conducive environment appears to be neonatal brain in which growth factors are readily available.

Animals

Biparietal diameter: a useful measure for determining gestational age of human abortuses.

BACKGROUND: The crown-rump length is conventionally used to determine the age of human abortuses. However, it is not reliable as it is dependent on the positioning of the conceptus. We compared this with the biparietal diameter and foot length for determining the gestational age. METHODS: Different measurements, commonly used to assess gestational age, were measured in 146 human abortuses for which an accurate obstetric history could not be elicited. Measurements taken were crown-rump length, biparietal diameter and foot length. These were correlated with the observations at antenatal examinations before finalizing the approximate age. RESULTS: Multiple regression analysis of the data indicated that of the three measurements, the biparietal diameter was the most reliable for determining foetal gestational age between 8 and 26 weeks. The age determined with the biparietal diameter correlated well with that of abortuses with an accurate obstetric history. CONCLUSION: The biparietal diameter of a human foetus may be used to determine its age if the obstetric history regarding the period of gestation is vague or not available.

Abortion, Induced