Creatine kinase in trout male germ cells: Purification, gene expression, and localization in the testis

Abstract

High creatine kinase (CK) activity (16.5 ± 7.6 IU/mg) is present in trout spermatozoa. In order to partly characterize the CK isozyme predominantly present in sperm and to study the expression of this protein in spermatogenesis, we purified to homogeneity a CK (s‐CK) from trout sperm, by nitrogen cavitation followed by two chromatography steps (DEAE‐Trisacryl and Blue Sepharose). Specific antisera to s‐CK were developed. A cDNA encoding for a CK named TCK1, and whose transcript shows enhanced testicular expression, was previously isolated from trout testis (Garber et al., 1990: Biochim Biophys Acta 1087:256–258). A CK subunit expressed in vitro by this cDNA cross‐reacts with anti‐s‐CK. A 21‐amino‐acid residue sequence near the N‐terminus of s‐CK is identical to the cDNA‐derived sequence of TCK1, which is unliked any previously reported CK sequence. Using in situ hybridization, the TCK1 mRNA was detectable in primary and secondary spermatocytes and in early spermatids. Immunohistochemical staining of testis and various organs revealed that s‐CK was confined to testis and, in this organ, to late spermatids and spermatozoa. In gill, some cells exhibited a positive signal, but another study rules out the presence of s‐CK in this organ (Garber et al., 1990: Biochim Biophys Acta 1087:256–258). These results demonstrate that s‐CK/TCK1 is a germ cell‐specific protein, the transcription of which starts in meiotic germ cells, while translation starts in late spermatids. © 1996 Wiley‐Liss, Inc.

Characterization of rainbow trout milt collected with a catheter: semen parameters and cryopreservation success

“…2). Presumably, it is the same protein as that described by Maisse, Pinson & Loir (1988) and later identi®ed as creatine kinase by Saudrais, Garber, McKay, Dixon & Loir (1996). These authors related low cryopreservation success of rainbow trout semen to the presence of this protein in seminal plasma.…”

Tissue and plasma enzyme activities and chemical analytes in Golden Trevally from a public aquarium

“…Creatine kinase expression in the gonads of Golden Trevally was highly variable and did not appear to be sex specific, which was not evaluated in either Black Sea Bass or Red Lionfish. However, CK activity has been described in the gonads of three species in the family Sciaenidae and was isolated from testes of Rainbow Trout and Green Sunfish Lepomis cyanellus (Fisher and Whitt 1978; Coppes et al 1987; Saudrais et al 1996). Creatine kinase activity is also stimulated by sex steroids in human and rodent models (Sömjen et al 1997; Katzburg et al 2001).…”

Acute Changes in Gill Na+‐K+‐ATPase and Creatine Kinase in Response to Salinity Changes in the Euryhaline Teleost, Tilapia (Oreochromis mossambicus)

“…Our Western data also showed the predominance of the MM-type CK in tilapia (Oreochromis mossambicus) gill after comparisons of MM-and BB-type CK. High CK activity is present in trout spermatozoa and is named as an s-CK isozyme (Saudrais et al 1996), but a previous study showed the absence of s-CK in gill (Garber et al 1990). Marine fish, red sea bream (Pagrus major), and Pacific mackerel (Scomber japonicus) possess less thermostable muscle CK than carp (Cyprinus carpio; Nakagawa and Nagayama 1991).…”