Iksookimia longicorpa and Cobitis hankugensis are two species of fish distributed only on the Korean Peninsula. They have a unique reproductive ecology that naturally hybridizes into three widely known unisexual types, maintaining populations of almost all females. In this study, the fine structure of the micropyles of I. longicorpa, C. hankugensis and their hybrids was analyzed to find out how egg–sperm interaction, a common interspecies isolation mechanism, is possible between heterogeneous species. Analysis of 30 eggs from five females of each species revealed that all had one funnel-shaped micropylar region and a manhole-shaped micropyle canal. With the exception of C. hankugensis, which had no spiral grooves or ridges, the rest had counterclockwise spiral grooves and ridges on the micropylar region. All five species, however, showed identical groove patterns for the micropyle canal. The egg size was the largest in HL (one from the C. hankugensis locus with one from the I. longicorpa locus) and the smallest in C. hankugensis. In the hybrids, the HL type had the largest egg and HHL (two from the C. hankugensis locus with one from the I. longicorpa locus) type the smallest. For the diameter of the micropylar region and micropyle canal, the diploid I. longicorpa, C. hankugensis and HL were smaller than those of the triploid. In addition, as the ratio of the canal diameter to the eggs was lower in I. longicorpa than in C. hankugensis, it was confirmed that I. longicorpa has a relatively small micropyle canal compared with C. hankugensis.

Structural studies in seeds with physical dormancy (PY) are important to better understand its causes and release when subjected to treatments for dormancy breaking. The aims of this study were to (1) characterize the PY break; (2) examine the role of different seed structures in water uptake; and (3) identify the water gap in Senna multijuga seeds. Imbibition patterns of dormant and non-dormant (subjected to dormancy breaking treatments) seeds and the morphological changes during dormancy breaking and germination were evaluated. To identify the water gap, the micropyle and lens were blocked separately, and the water absorption by seed parts was determined. Structural characteristics of the seed coat were also examined. Immersion in water at 80°C was efficient in breaking seed dormancy and imbibition occurred first at the hilar region, through the lens. Water was not absorbed through the micropyle or the extra-hilar region. S. multijuga seeds have a testa with a linearly aligned micropyle, hilum and lens. The seed coat consisted of a cuticle, macrosclereids, one (hilar region) or two (extra-hilar region) layer(s) of osteosclereids and parenchyma cell layers. The lens has typical parenchyma cells underneath it and two fragile regions comprised of shorter macrosclereids. Heat treatment stimulated the lens region, resulting in the opening of fragile regions at the lens, allowing water to enter the seeds. It is concluded that short-term exposure to a hot water treatment is sufficient for the formation of a water gap in S. multijuga seeds, and only the lens acts in the imbibition process.

In the present work, we analyzed the structure of oocytes and fertilized eggs of the piracanjuba fish (Brycon orbignyanus) under light and scanning electron microscopy. After inducing spawning, samples were collected at the moment of oocyte extrusion, when oocytes and semen were mixed (time 0), as well as at 10, 20 and 30 s after mixing, every minute up to 10 min, and then at 15 and 20 min. The oocytes are spherical, translucent and greenish with a mean diameter of 1.3 ± 0.11 mm. During the extrusion, cytoplasmic movement was observed in eggs towards the micropyle, characterizing the animal pole. At the moment of fertilization, the cortical cytoplasm showed a higher concentration of cortical alveoli at the animal pole than at the vegetal pole. The cortical alveoli breakdown promoted the elevation of the chorion with a consequent increase in egg diameter (1.95 ± 0.08 mm). The penetration of the spermatozoon promotes the formation of a fertilization cone of spherical external structure, which obstructs the opening of the micropyle. This structure acts as a main mechanism to avoid polyspermy, intercepting the access of supernumerary spermatozoa. Such studies about the reproductive biology of fish are important to species survival and conservation programmes.

Scanning electron microscopic examination of the egg shell and associated structures of Eocanthecona furcellata (Wolff.) (Hemiptera: Pentatomidae) reveals a distinct pattern of structural elements, viz. microsculpture of the chorion, pseudoperculum, egg burster and aeromicropylar processes. The egg shell has a highly decorated chorion with a structural difference at both the anterior and posterior ends. A reticulate type of microsculpture with varying sizes of spine-like projections measures 4.02 ± 0.06 to 8.12 ± 0.05 μm and forms an asteroid-like shape at the anterior pole, while the surface at the posterior end has elongated striations over the chorion measuring 72.00 ± 0.074 μm. The pseudoperculum of the egg shell is a circular lid measuring 518 ± 11.43 μm in diameter. During hatching the line of rupture follows the micropylar ring, and this sometimes extends beyond the ring of micropylar processes. An inverted T-shaped black egg burster measures 202 ± 4.63 μm in length, is well developed and can be seen prior to hatching within a thin and transparent egg shell. It is a median sclerotised part of embryonic cuticle. The aeromicropylar processes are situated at the anterior pole of the barrel-shaped eggs. There are 26–28 aeromicropylar processes arranged in a circle at almost equidistance and each micropylar process is slightly curved opposite to the pseudoperculum lid with a bulbous end at the apex. The aeromicropylar processes are 25.40 ± 0.083 μm in length and 3.02 ± 0.07 μm in width. Each micropylar process has a central canal for the passage of sperm whereas the rest of its body is porous, and allows respiratory interchange.

The process of sperm penetration into the egg of the European catfish Silurus glanis L. was investigated by scanning and transmission electron microscopy. Unfertilized ovum membrane under the internal micropylar aperture shows a critical depression with undulation and short microvilli corresponding to a specialized sperm entry site structure. Twenty seconds after insemination, the fertilizing spermatozoon fused with the egg at this specialized structure, without formation of an earlier fertilization cone, which had been confirmed in eggs of other fish. A cytoplasmic eminence developed however, immediately under the fused sperm head and then disappeared with time after fertilization to form a circular area around the fused sperm head, which in turn became embedded in the circular area with shortening of the flagellum. The micropyle in the catfish egg consisted of a micropylar vestibule and a canal, the internal micropylar canal being wide enough to permit the penetration of only one spermatozoon.