Cloning and characterization of the rat Crisp-1 gene.

Rat androgen-regulated acidic epididymal glycoprotein (AEG), also known as Protein DE, is a product of the Crisp-1 gene. Protein DE is secreted into the epididymal lumen and binds to sperm heads during their transit through the epididymis. In experiments reported here, the rat Crisp-1 gene has been cloned and its structure determined.

Identification of osteopontin (OPN) mRNA and protein in the rat testis and epididymis, and on sperm.

We have identified a bone cell adhesion molecule, osteopontin, in the rat testis and epididymis by Northern analysis, RT-PCR, Western immunoblot analysis and immunocytochemistry. A polyclonal antibody raised against rat epididymal fluid proteins was used to detect fusion proteins produced by a testis lambda gt11 cDNA library. Sequence analysis of one of four positive cDNA clones, designated as pREP5, revealed identity with the rat osteopontin (OPN) cDNA.

Testicular synchrony: evaluation and analysis of different protocols.

Using the vitamin A depletion-replacement rat model to obtain testicular synchrony, we examined the reproducibility and degree of synchronization obtained by two different protocols. In the original protocol (A), synchrony was achieved by use of retinol alone. In protocol B, retinoic acid was used during the final days of vitamin A depletion as a supplement to retinol.

Function of vitamin A in normal and synchronized seminiferous tubules.

Vitamin A is clearly an important factor in spermatogenesis. Some of the new data on metabolism of retinoids in the testis has contributed to our understanding of the mechanism(s) involved in the action of vitamin A. It is probable that the requirement of the testis of vitamin A deficient rats for retinol but not retinoic acid involves access of the retinoids to various testicular compartments.

Partial characterization of a fraction from human follicular fluid that initiates the human sperm acrosome reaction in vitro.

A human follicular fluid (HFF) fraction prepared by Sephadex G-75 column chromatography has been previously shown by this laboratory to initiate the human sperm acrosome reaction (AR) in vitro. In the present report, the apparent molecular weight (MW) of this AR activity determined by a longer G-75 column than was used in the previous work was 50,000 +/- 5,106. The G-75 Sephadex void volume fractions of some but not all HFF samples were also found to contain some AR-initiating activity.

Human sperm acrosome reaction-initiating activity associated with the human cumulus oophorus and mural granulosa cells.

This report describes the detection and partial characterization of preovulatory human cumulus oophorus and mural granulosa cell-associated activity capable of initiating the human sperm acrosome reaction (AR) in vitro. Fragments of preovulatory human cumulus (cells plus extracellular matrix) were washed 3 times, incubated for 24 hr and the spent media and washes assayed for their ability to initiate the human sperm acrosome reaction (AR) in vitro. AR activity was present in the first two washes but not the third wash; however, AR activity was recovered in the spent medium after 3 X-washed fragments were incubated for 24 hr under conditions which maintained the viability of the cumulus cells.

Hamster sperm Na+, K+-adenosine triphosphatase: increased activity during capacitation in vitro and its relationship to cyclic nucleotides.

These in vitro studies of golden hamster sperm were undertaken to determine whether: Na+, K+-adenosine triphosphatase (ATPase) activity is required for capacitation; Na+, K+-ATPase activity is altered during capacitation; and cyclic nucleotides can control this enzyme activity. Hamster sperm were incubated in a medium in which capacitation occurred in an asynchronous manner and in which acrosome reactions began to occur after approximately 3.5 h of incubation.

Immunologic and biological properties and clinical significance of placental proteins PP5 and PP12.

Among the number of newly isolated placental proteins, PP5 and PP12 share some common characteristics: Both are present in the syncytiotrophoblast of normal placenta and hydatidiform mole, but less frequently, if at all, in choriocarcinoma. The levels in heparinized plasma of both proteins are lower than those in serum, and both are heat-labile. The function of PP12 is completely unknown, whereas PP5 appears to be related to the blood coagulation and fibrinolytic systems at the placental site through its antiplasmin activity.

Placental protein 5 is related to blood coagulation and fibrinolytic systems.

Previous studies have shown that placental protein 5 (PP5) forms complexes with heparin. In order to further elucidate the biological role of PP5 we studied the effect of plasmin and thrombin on the immunoreactivity of PP5, and the possible functional antiplasmin and antithrombin effects of purified PP5. Varying concentrations of plasmin and thrombin were added to pregnancy plasma, and the PP5 levels, measured by radioimmunoassay, were found to be elevated by 558% (plasmin and 48-87% (thrombin).

Placental protein 5 (PP5)-like immunoreactivity in human urine.

Placental protein 5 (PP5)-like immunoreactive material was detected in human male urine in small concentrations (17-60 pg/ml). Part of the PP5 immunoreactivity in the urine had a molecular weight of 36 000-42 500, which is the molecular weight of purified PP5 from the human placenta. In radioimmunoassay, serial dilutions of the 36 000-42 500 molecular weight material gave an inhibition curve parallel to that of the PP5 standard.

Human seminal plasma contains a protein that shares physicochemical and immunochemical properties with placental protein 5 from the human placenta.

Placental protein 5 (PP5)-like immunoreactive material was detected in human seminal plasma at high concentrations (32-1000 ng/ml). This was not due to interference with proteases or binding to seminal plasma proteins, since immunoreactivity was not affected by treatment with protease inhibitors, and incubation with seminal plasma of [125I]PP5 did not bring about any significant change in the elution pattern in gel filtration. Part of the PP5 immunoreactivity in seminal plasma had a molecular weight of 36,000-42,500 which is the molecular weight of purified PP5 from the human placenta.