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Biomedical subjects

M Sousa

Publications and source records attributed to M Sousa.

62 records · Page 4Linked to original sources

Silver staining of the postacrosomal sheath during human spermiogenesis.

Applying the silver staining technique, it could be shown that in the early phase of spermiogenesis a layer of argyrophilic material accumulated at the base of the acrosomal vesicle and at the outer side of the nuclear envelope opposite that region, and, later, at the inner side of the plasma membrane near the base of the acrosomal vesicle. During further development of the postacrosomal region of the spermatozoon head, the argyrophilic material associated with the plasmalemma grew caudally to form the postacrosomal dense lamina, while the argyrophilic material associated with the nuclear envelope, staying the same size, shifted to the caudal end of the postacrosomal dense lamina to form the post-nuclear band.

Humans↗

Ultrastructural and cytochemical study of spermatogenesis in Scrobicularia plana (Mollusca, Bivalvia).

The ultrastructure of the mature spermatozoa and spermatogenesis of the bivalve Scrobicularia plana are described. Support cells extend from the basal lamina to the lumen of the testis and are laterally connected to the germinal epithelium. Germ cells present intercellular bridges and flagella since the spermatogonial stage. While spermatogonia and spermatocytes appear connected to support cells by desmosome-like junctions, elongated spermatids are held at the acrosomal region by support cell finger-like processes. During spermiogenesis, the acrosomal vesicle differentiates from a golgian saccule and then migrates to the nuclear apex. A microtubular manchette arising from centrioles surrounds the acrosomal vesicle, the nucleus, and the mitochondria at the time these three organelles start their elongation, disappearing after that. The mature spermatozoon of S. plana lacks a distinct midpiece because the mitochondria extend from the region of the pericentriolar complex along the nucleus anteriorly for approximately 1.4 microns. The features of this bivalve type of modified spermatozoon are compared with those of other animal groups having similar modifications.

Acid Phosphatase↗

Starfish sperm-oocyte jelly binding triggers functional changes in cortical granules. A study using acid phosphatase and ruthenium red ultrastructural histochemistry.

Starfish oocytes were examined before fertilization, immediately after insemination, and during the cortical reaction by means of acid phosphatase and ruthenium red ultrastructural histochemistry. Oocyte cortical granules are composed of a lamellar body and a surrounding matrix which is subdivided into dense and light portions. In unfertilized oocytes cortical granules are not stained by ruthenium red but show a weak acid phosphatase activity in the light portion of the granule matrix. Immediately after the adhesion of the spermatozoon to the oocyte jelly coat, the light matrix portion of cortical granules appears stained by ruthenium red and shows a strong acid phosphatase activity. During the cortical reaction, cortical granules are released into the perivitelline space and the lamellar body, surrounded by the stained matrix, fuses with the fertilization envelope. Our data suggest that membrane permeability changes and enzyme activation occur in the egg when the spermatozoon binds to the oocyte jelly coat.

Acid Phosphatase↗

Presence of ATPase and alkaline phosphatase activities in the starfish sperm acrosome.

ATPase activity was cytochemically detected in the peripheral acrosomal component of ionophore-reacted sperm, while alkaline phosphatase activity was demonstrated in the upper and central components of the acrosome and, at fertilization, at the site of sperm-oocyte binding. Supernatants of ionophore treated sperm suspensions were assayed for ATPase, alkaline and acid phosphatase activities. Results suggest that alkaline phosphatase may be involved both in the acrosomal reaction and oocyte jelly lysis but the function of the acrosomal ATPase remains unknown.

Acrosome↗

Starfish acrosomal acid phosphatase: a cytochemical and biochemical study.

The acrosome reaction of spermatozoa from the starfish Marthasterias glacialis was induced with the ionophore A23187. Reacted cells were then processed for acid phosphatase ultrastructural cytochemistry, but significant enzyme activity was not detected. However, when the supernates from suspensions of ionophore-treated sperm were assayed for acid phosphatase, a net enzyme activity was observed. Supernatant proteins were run in starch gel electrophoresis and fluorescent zymograms revealed a single band of acid phosphatase. SDS-PAGE of proteins eluted from the active spots of starch gels showed one major band of about 63 kDa. The results obtained support the hypothesis that the acid phosphatase whose activity has been detected only at the time of binding of sperm and egg originates from the sperm acrosome.

Acid Phosphatase↗

Birth after electroejaculation coupled to intracytoplasmic sperm injection in a gun-shot spinal cord-injured man.

We describe the case of a man who, after a gun-shot wound, has become paraplegic. As a consequence of his spinal cord injury he developed infertility due to incomplete erection and anejaculation. After several unsuccessful penile vibratory stimulation attempts, it was possible to achieve ejaculation by means of transrectal electrostimulation. However, the total sperm motility count, the percentage of normal sperm morphology, and the vitality and hypo-osmotic swelling test scores were rather low, and in particular viability was strikingly decreased during sperm preparation. These parameters further decreased following electroejaculate, forcing us to use intracytoplasmic sperm injection instead of conventional in vitro fertilization for the treatment cycle. A normal pregnancy was achieved which resulted in birth of a healthy girl on 1st January 1997.

Adult↗

Fine silver staining analysis of the nucleolar organizer regions during oogenesis in Penaeus kerathurus (Crustacea, Decapoda).

In previtellogenic oocytes, nucleoli showed segregated components, with the dense fibrillar component (DFC) appearing highly developed and presenting several fibrillar centers (FC). The granular component (GC) was less developed and formed a wide-spaced reticulum. Only the DFC appeared stained by silver, with higher intensities being found at its periphery. During early vitellogenesis, the nucleolar components were kept segregated but both the DFC and the GC enlarged, without evident changes being noticed in the silver staining pattern. In mid and late vitellogenesis, the nucleoli showed integrated components, with the DFC being intermeshed with the GC. Both nucleolar components were highly developed, no evident FC were noticed, and silver stained the DFC in a heterogeneous pattern. During cortical vesicle formation, the nuclear chromatin condensed and nucleoli appeared disintegrated, showing high levels of accelerated exportation of silver stained materials. Results suggest that the size of the DFC is kept high and the size of the GC kept low (low rDNA transcription levels and RNP exportation accelerated), in the segregated nucleoli of the previtellogenic oocyte, as the cell stores nuages but shows absence of rough endoplasmic reticulum and thus low protein svnthesis; that the size of the DFC and of the GC is increased in the segregated nucleoli of early vitellogenic oocytes (intermediate levels of rDNA transcription and of protein synthesis), which is in accordance with the appearance of the rough endoplasmic reticulum and of yolk vesicles formed with endogenous and exogenous sources; that during mid and late vitellogenesis the DFC and the GC appear highly developed and integrated (high levels of rDNA transcription and of protein synthesis) as the rough endoplasmic reticulum expands and the large yolk vesicles grow by endogenous synthesis; and that chromatin condense and nucleoli disintegrate (very low levels of rDNA transcription with accelerated RNP exportation) when cortical vesicles are formed.

Animals↗