Sperm Heads

The Starting material used for preparing both nucleoprotamines and protamines is sperm heads or testis chromatin of fish obtained from freshly collected mature milts or sperm freshly drawn from mature testes. These mature milts or sperm contain essentially only one type of cell, spermatozoa, although the former often contain immature sperm cells and connective tissues which must be removed. Ways of preparing sperm heads, chromatin, nucleoprotamines, and protamines will be briefly reviewed in this chapter  

The preparation of sperm heads from fish is fairly easy, because the spermatozoa of fish are apt to lose their tail fibrils on stirring or homogenizing in an appropriate aqueous solution, leaving the sperm heads intact. With progress in methods of isolating cell nuclei, several modifications have been added to the original procedure of Kossel (1929). 

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Pagulayan IF, Baoanan ZG, Villa LQ. 1994. The effect of methyl parathion on the sperm head morphology of the ICR strain mice. Asia Life Sci 3(1) 45-54. 

Rat (Sprague- Dawley) 1 d 2x (G) 20000 M (transient decreased sperm head count per gram testis) Linder et al. 1992... 

Fig. 41.1 Micrographs of boar spermatozoa treated with Allium extraets (0.5 g/mL) (a) garlic bulb extract, (b) untreated spermatozoa. Formation of a clear halo around the sperm head indieated acrosin proteolytie activity...

Topham JC. 1980. Do induced sperm-head abnormalities in mice specifically identify mammalian mutagens rather than carcinogens Mutat Res 74 379-387. 

Fig. 3. (A) Stereo pair of starfish sperm chromatin fibers stained with osmium ammine-B. Bracket indicates the fine deposition of stain on nucleosomes and linker DNA. Arrowheads indicate where fibers enter and exit the plane of the section, and asterisks indicate sharp bends in the fibre axis. (B) Stereo pair of a reconstructed volume of a starfish sperm head by EM tomography. Axes of some fibers have been marked. Arrowhead indicates where individual fibers cannot be distinguished (from Ref. [27]). Scale bar 100 nm.

Both positive and negative results have been reported in in vitro genotoxicity smdies of chlorine dioxide." In vivo assays did not find increases in micronucleus induction, chromosomal aberrations, or sperm head morphology after oral exposure, but they did find increases in micronucleus induction after intraperitoneal injection."... 

Oral administration of lOOOmg/kg by gavage to male rats for a total of 12 treatments over 16 days caused reduced testes weight and increases in abnormal sperm heads. ... 

Meier JR, Bull RJ, Stober JA, et al. 1985. Evaluation of chemicals used for drinking water disinfection for production of chromosomal damage and sperm-head abnormalities in mice. Environ Mutagen 7 201-211. 

AOT/isooctane Protamines from sperm head nucleoproteins of fish Conformational studies on model polypeptides with amino acid sequences similar to clupeine and salmine were carried out in RMs [286]... 

Thorgeirssorr, S.S., Glowirrski, LB. McManus, M.E. (1983) Metabolism, mutagerricity and carcinogerricity of aromatic amines. Rev. Biochem. Toxicol, 5, 349-387 Topham, J.C. (1980) Do induced sperm-head abnormahties in mice specifically iderrtily mammalian mutagens rather than carcinogerrs Mutat Res., 74, 379-387... 

Fertilization is the fusion of the sperm and ovum. The sperm head binds to the plasma membrane of the egg (oolemma), and the entire spermatozoon enters the cytoplasm of the ovum. Only capacitated spermatozoa with intact acrosomes can enter and pass through the cumulus oophorus. The acrosome is a membrane-bound, cap-like structure covering the anterior portion of the sperm nucleus. The acrosomal reaction is the release of materials that lyse the glycoprotein coat (zona pellucida) surrounding the ovum. This is necessary for fertilization to take place. Before undergoing the acrosomal reaction, sperm go through a type of hypermotility called hyperactivation. The... 

The mass of bull sperm heads in a sample was determined by interference microscopy, and the following results were obtained ... 

Butadiene increased the frequency of sister chromatid exchanges and micronuclei in mouse but not rat bone marrow. Micronucleus frequency also increased in peripheral erythrocytes and splenocytes. Butadiene also induced chromosomal aberrations in mouse bone marrow, and dominant lethal mutations, heritable translocations and sperm-head abnormalities in mice. It did not induce aneuploidy in bone marrow cells in vivo. 

In a single study, epichlorohydrin bound to DNA of mice and rats treated in vivo. One study reported that sister chromatid exchanges were induced in the bone marrow of partially hepatectomized CBA/J mice treated with epichlorohydrin by a single intraperitoneal injection. Sister chromatid exchange frequencies in mice that did not receive partial hepatectomy before treatment with epichlorohydrin were comparable to the control frequencies. One of two studies reported that epichlorohydrin induced chromosomal aberrations in mouse bone marrow. Positive results were also reported for epichlorohydrin in the mouse host-mediated assay in one of three studies. In single studies, epichlorohydrin caused sperm head abnormalities in rats but not mice. It did not induce micronuclei or dominant lethal mutations in mice in vivo. 

Cassidy, S.L., Dix, K.M. Jenkins, T. (1983) Evaluation of a testicular sperm head counting technique using rats exposed to dimethoxyethyl phthalate (DMEP), glycerol alpha-mono-chlorohydrin (GMCH), epichlorohydrin (ECH), formaldehyde (FA), or methyl methane-sulfonate (MMS). Arch. Toxicol., 53, 71-78... 

Toluene did not induce sperm-head abnormalities in mice. 

Trichloroethane covalently bound to DNA, RNA and protein in mice and rats but did not induce micronuclei or abnormal sperm head morphology in mice in vivo. It induced chromosomal aberrations and cell transfonnation in mammalian cell cultures and it showed inconclusive evidence of sister chromatid exchange induction. It did not induce unscheduled DNA synthesis or gene mutation in mammalian cells in vitro. 

Topham, J.C. (1980) Do indueed sperm-head abnormalities in miee speeifieally identify mammalian mutagens rather than eareinogens Mutat. Res., 74, 379-387... 

Meier, J.R., Bull, R.J., Stober, J.A. Cimino, M.C. (1985) Evaluation of ehemieals used for drinking water disinfeetion for produetion of ehromosomal damage and sperm-head abnormalities in miee. Environ. Mutag., 7, 201-211... 

I The first and simplest of them are those of sperm-head nucjeo-protein [17]. Here the nucleic acid, DNA, seems to determine the structure. In the gap already mentioned between the coils of the double spiral there is no room for any more nucleotide, but only for the smaller protamine chain, as Wilkins has shown. No similar structure has been found for natural RNA but it does occur for the combination of two different artificial poly-adenosine ribose phosphate and guanine ribose phosphate. This fact, established by Rich [18], is the best example of heterogeneous mutual coiling in vitro of polymer molecules. 

In 1868 Friederich Miescher isolated a substance from the nucleus of pus cells, to be characteristic of the nucleus, and he called it nuclein. A similar substance isolated from salmon sperm heads. Nuclein was later shown to be a mixture of a... 

Sheep and goat milk, palm seed oils, sperm head oil... 

Mouse (spermatogonia) Sperm head abnormality + Topham1980... 

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