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Biomedical subjects

F J Longo

Publications and source records attributed to F J Longo.

At least 73 records · Page 4Linked to original sources
PubMed1979

Differences in mouse ovarian cells as distinguished by horseradish peroxidase labelling.

The extracellular space of mouse ovarian follicles and stroma contained horseradish peroxidase (HRP) reaction product one minute after intravenous injection of the tracer. In addition to pinocytotic uptake of the tracer, non-vesicular staining with HRP reaction product, heretofore unrecognized, was noted in a variety of ovarian cell-types. This diffuse staining was correlated with changes in cellular morphology suggestive of degeneration. These findings are discussed in relation to the composition of cell populations comprising follicles and ovarian stroma and alterations in cell function.

Animals↗
PubMed1979

Cytological events associated with in vitro aged and fertilized rabbit eggs.

The morphogenetic events associated with rabbit eggs aged in vitro for 12 to 50 hours prior to mixing with sperm have been examined by light and electron microscopy. After 12 hours in culture, morphological alterations of the meiotic spindle and the cortex of unfertilized eggs were evident. By 24 to 50 hours in culture, unfertilized eggs contained subnuclei, structures which formed when individual and/or groups of meiotic chromosomes dispersed and becmae invested by a double-laminated structure reminiscent of a nuclear envelope. Although most eggs obtained 11.5 to 12 hours after induced ovulation and in vitro fertilized displayed morphogenetic patterns similar to those described for in vivo fertilized ova, some (10%) contained three pronuclei. Many eggs obtained 13 to 15 hours after induced ovulation and subsequently mixed with sperm in vitro appeared to undergo processes of fertilization typical of in vivo fertilized eggs, however, approximately 30% contained subnuclei in association with the male pronucleus. Few eggs (15%) aged 12 hours prior to in vitro fertilization displayed patterns of pronuclear development and association typical of fertilized unaged ova. Subnuclei developed in many of the fertilized ova. Supernumerary sperm nuclei, which did not develop into male pronuclei, were observed in some zygotes. Cleavage of eggs aged 12 hours prior to fertilization was abnormal or retarded. After 24 hours in culture approximately 16% of the eggs fertilized. Seventy percent of the fertilized eggs failed to support the development of a male or female pronucleus.

Animals↗
PubMed1978

Effects of puromycin and actinomycin D on fertilized rabbit eggs cultured in vitro.

Rabbit zygotes were cultured in media containing varying concentrations of puromycin and actinomycin D in order to determine what effect these agents have on events of fertilization and early development. Concentrations of puromycin greater than 10(-5) M inhibited mitosis, cytokinesis of fertilized eggs and leucine incorporation. In approximately 50% of the zygotes treated with puromycin (10(-5) M or greater) the female pronucleus remained at its site of formation, the cortex, and did not migrate centrad. In these instances the male pronucleus moved from its site of development to the female pronucleus. Concentrations of actinomycin D greater than 10(-7) M also inhibited mitosis and cytokinesis of fertilized eggs. Breakdown of the nuclear envelopes and association of the paternally- and maternally-derived chromatin did not occur in either puromycin- or actinomycin D-treated embryos. Actinomycin D at all of the concentrations employed (10(-9) to 10(-4) M) had little effect upon the structure of nucleolus-like bodies within male and female pronuclei. Male and female pronuclei of puromycin (10(-4) M) and actinomycin D (10(-5) M) treated zygotes, incubated with tritiated thymidine and prepared for scintillation counting and autoradiography, synthesized DNA at a reduced level when compared to control specimens. Although "side effects" of puromycin and actinomycin D treatment are not excluded, these results suggest that nascent proteins may be required for events of fertilization.

Animals↗
PubMed1978

Nuclear protein changes in the maternally and paternally derived chromatin at fertilization.

The proteins which become associated with the paternally derived chromatin during fertilization may be instrumental in its activation and in the dramatic structural metamorphosis of the sperm nucleus during pronuclear development. Proteins associated with sperm and zygote nuclei and male and female pronuclei of fertilized sea urchin eggs were analysed by polyacrylamide gel electrophoresis in order to examine nuclear protein changes in the paternally and maternally derived chromatin following insemination. Results demonstrate major changes in both the solubility characteristics and polypeptide profiles of sperm nuclei upon insemination. Evidence is presented which indicates that at fertilization the paternally derived chromatin acquires proteins of molecular weights greater than 80,000 and a nuclear protein composition similar to that of the female pronucleus. The nuclear proteins associated with zygote nuclei were compared to those of combined male and female pronuclei and showed many similarities and some differences. Several polypeptides were present in zygote nuclei which were not observed in pronuclear extracts.

Animals↗
PubMed1977

Synthesis of RNA by male pronuclei of fertilized sea urchin eggs.

Experiments have been conducted in an effort to determine when the paternally derived genome of sea urchin embryos becomes active in RNA synthesis. Participation of the paternal genome in RNA transcription as early as fertilization has been indicated by the following: (1) polyspermic zygotes consistently demonstrate greater levels of incorporation of labelled precursor (3H-uridine) into RNA when compared to monospermic eggs. (2) Monospermic and polyspermic eggs treated with actinomycin D show the same levels of incorporation of label into RNA. (3) Analysis of autoradiographs demonstrates the incorporation of 3H-uridine into male and female pronuclei.

Animals↗
PubMed1977

An ultrastructural study of cross-fertilization (Arbacia female x Mytilus male).

Insemination of sea urchin (Arbacia) ova with mussel (Mytilus) sperm has been accomplished by treating eggs with trypsin and suspending the gametes in seawater made alkaline with NaOH. Not all inseminated eggs undergo a cortical granule reaction. Some eggs either elevate what remains of their vitelline layer or demonstrate no cortical modification whatsoever. After its incorporation into the egg, the nucleus of Mytilus sperm undergoes changes which eventually give rise to the formation of a male pronucleus. Concomitant with these transformations, a sperm aster may develop in association with the centrioles brought into the egg with the spermatozoon. Both the male pronucleus and the sperm aster may then migrate centrad to the female pronucleus. Evidence is presented which suggests that fusion of the male pronuclei from Mytilus sperm with female pronuclei from Arbacia eggs may occur, although this was not directly observed. These results demonstrate that Mytilus sperm nuclei are able to react to conditions within Arbacia eggs and differentiate into male pronuclei.

Acrosome↗
PubMed1976

Sperm aster in rabbit zygotes: its structure and function.

Microscope observations of rabbit zygotes demonstrate that a sperm aster forms in association with the male pronucleus approximately 1 h postinsemination and consists of two regions. One, the centrosphere, contains a dense aggregation of cisternae of smooth endoplasmic reticulum and microtubules. The second consists of fascicles of microtubules which emanate from the centrosphere. Fertilized rabbit eggs were cultured in medium containing colcemid in order to determine its effects on various events of fertilization, such as movements of the male and female pronuclei and DNA synthesis. No evidence was obtained to indicate that a sperm aster is formed in colcemid-treated zygotes. In addition, migration and close apposition of the pronuclei do not take place. Breakdown of the pronuclear envelopes and condensation of the maternally and paternally derived chromosomes occur even though the pronuclei fail to migrate centrad. Autoradiographic analysis of the synthesis of DNA by both pronuclei demonstrates that their migration into close apposition to one another is not required for the incorporation of tritiated thymidine.

Animals↗
PubMed1975

Ultrastructural analysis of artificially activated rabbit eggs.

Ultrastructural investigations have been carried out on parthenogenetic rabbit eggs in an effort to elucidate events occurring during artificial activation and their similarity to processes of fertilization and embryogenesis. Rabbit eggs were artificially activated by culturing at 10 degrees C for 24 hours followed by incubation at 37 degrees C for 2 to 24 hours. Examination of eggs immediately after incubation at 10 degrees C for 24 hours indicates that activation is initiated when the chromosomes coalesce to form a reticulum which is either surrounded completely by two parallel membranes or incompletely by cisternae of smooth endoplasmic reticulum. Aggregation of the chromosomes occurs as a result of a reduction in the number of microtubules making up the meiotic spindle. When cold treated ova are subsequently incubated at 37 degrees C a nucleus is formed which moves central where it may participate in the cleavage of the egg. Formation of a second polar body and release of the contents of the cortical granules as reported for inseminated eggs was not found to be a part of activation of the egg by cold treatment. Approximately 95% of the ova cultured at 10 degrees C for 24 hours followed by 37 degrees C for 12 hours were activated, i.e., they possessed a nucleus or they had cleaved. Many of the activated eggs cultured for short periods at 37 degrees C were structurally similar to fertilized ova, with further incubation fragmented eggs and abnormal multicellular stages predominated.

Animals↗
PubMed1975

Cytological events leading to the cleavage of golden hamster zygotes.

Fertilized golden hamster eggs were examined between 6 and 20 hours post-ovulation to determine the events leading to the two-cell stage. Following their migration the pronuclei remain in the central region of the zygote for approximately ten hours. The morphologically, indistinguishable male and female pronuclei remain relatively unchanged during this period, i.e., they do not interdigitate or fuse with one another as described for the zygotes of other organisms. Following this period and at the time of pronuclear breakdown elongate vesicles appear along the nucleoplasmic surface of the pronuclear envelopes. Later the pronuclear envelopes fragment into elongate cisternae; these and the vesicles formed along the inner lamina of the pronuclear envelopes remain closely associated and constitute quadrilaminar structures. The chromosomes which condense prior to and during pronuclear envelope breakdown, migrate to the equatorial plate of the forming cleavage spindle. After cytokinesis the chromosomes in the blastomere nuclei disperse. Increase in the nuclear envelope to accommodate this dispersion may involve the addition of membrane from the quandrilaminar structures.

Animals↗
PubMed1975

Ultrastructural analysis of the testes in male pseudohermaphrodism reductase.

Light and electron microscopic observations of the testes in male pseudohermaphrodism due to deficiency of 17-ketosteroid reductase demonstrate an increased thickness of the walls of the seminiferous tubules. Although the seminiferous tubules are filled mostly by Sertoli cells containing crystalloids of Charcot-Bottcher, clusters of spermatogonia and spermatocytes are located at infrequent intervals along their lengths. Differentiation of spermatogenetic cells beyond the spermatocyte stage was not observed. Hyperplasia of Leydig cells, which are structurally similar to those of the normal testis, was pronounced. Pigment bodies were present in Leydig cells, whereas crystals of Reinke were not observed. The decreases in plasma androstenedione, testosterone and estrone following orchiectomy and the presence of a well-developed system of organelles (smooth endoplasmic reticulum and mitochondria containing tubular cristae), typical of steroid-secreting cells, indicate that the Leydig cells were active in steroid hormone synthesis, albeit deficient in 17-ketosteroid reductase activity.

17-Ketosteroids↗