Identification of unstimulated constitutive immunocytes, by enzyme histochemistry, in the coenenchyme of the octocoral Swiftia exserta.

Most animals rely on circulating hemocytes as cellular effectors of immunity. These cells traditionally have been characterized by morphology, function, and cellular contents. Morphological descriptions use granule differences and cell shapes; functional descriptions rely on phagocytic ability and oxygen transport; and cellular content descriptions include cytochemical features and key enzymes. Key enzymes used to identify phagocytes in tissues include hydrolytic enzymes, peroxidase, and--in invertebrates--phenoloxidase. Cnidaria such as Swiftia exserta lack a circulatory system, thereby complicating the identification of immune effector cells. As a first step in identifying immunocytes, this study focused on basic enzymes used during phagocytosis and encapsulation; both processes have been reported in octocorals such as S. exserta. Earlier work suggested that there are two populations of phagocytic cells: a constitutive population and an induced population following a trauma-associated challenge. To identify the constitutive immune effector cells in S. exserta in a nonactivated state, we used cryosections of unstimulated animals and the following enzymes to serve as identifying proxies due to their roles in phagocytosis and encapsulation: (1) acid phosphatase, (2) alkaline phosphatase, (3) non-specific esterase, (4) ß-glucuronidase, (5) peroxidase, and (6) phenoloxidase. Our results indicate that in unstimulated animals, two distinct cell populations could function as immunocytes. These cell types were differentiated by their enzyme reactivity and their location within the mesoglea of S. exserta, and have been described as either "oblong granular cells" or "granular amoebocytes."

Characterization and phylogenetic analysis of a cnidarian LMP X-like cDNA.

Proteasomes are multisubunit protease complexes which are partly responsible for metabolism of intracellular, ubiquitinylated proteins. Vertebrates have adapted a second and specialized structure responsible for the generation of peptides presented to the adaptive immune system and is thus, commonly referred to as the immunoproteasome. This complex is assembled from paralogous copies of subunits belonging to the constitutive, housekeeping proteasome. The immunoproteasome is more efficient in the generation of peptides for display on major histocompatibility complex (MHC) molecules. Important components of this complex are the paralogous members, LMP X and 7; where the latter replaces the former in the assembly of the immunoproteasome of vertebrates. In this report, we describe an LMP X-like cDNA from an endosymbiont-free gorgonian coral, Swiftia exserta. Cnidarians predate the phylogenetic divergence of protostomes and deuterostomes (P-D split), and are becoming an essential model for our comprehension of immune system evolution. Phylogenetic analyses of available sequences indicates that invertebrate LMP X-like sequences are outgroups to vertebrate LMP X and LMP 7, and is in agreement with previous observations that the duplication event giving rise to the two rapidly diverging lineages of proteasomal subunits occurred before jawed fished divergence.

Characterization of a C3-like cDNA in a coral: phylogenetic implications.

C3, C4, and C5 are thiolester-containing proteins (TEPs) of vertebrate complement. The identification of the molecular origin of the TEP family, and more specifically the ancestor protein of complement components C3, C4, and C5, remains a fundamental question. The prevailing paradigm suggests that duplication and divergence of these proteins occurred after the deuterostome split in phylogeny. It is believed that the ancestor of thiolester-containing complement proteins was alpha-2-macroglobulin (A2M)-like, a noncomplement-related protein. Here we describe a C3-like cDNA from a gorgonian coral, Swiftia exserta. This study provides evidence for the origins of a complement-related C3-like gene in the Precambrian period, predating both protostomes and deuterostomes. Furthermore, one may speculate that complement-like opsonic reactions were evolving at the earliest stages of metazoan evolution. This calls for a reassessment of the present concepts concerning the origins and evolution of TEPs.

The cellular basis of the aggressive acrorhagial response of sea anemones.

Some sea anemones possess structures called acrorhagi at the base of the tentacles. The acrorhagi are utilized solely for aggression. Acrorhagial aggression involves very exquisite intra- and interspecific recognition. This study examined acrorhagi and putative acrorhagial analogues or homologues in four species of sea anemone. The morphology and ultrastructure of tentacles, pseudoacrorhagi, column vesicles, and verrucae (adhesive column vesicles) differed from that of acrorhagi. Coral capitate tentacles and acrorhagi have different surface morphology, nematocysts, and functions. Besed on morphology, acrorhagi seem more likely to be homologous to tentacles than to verrucae. Acrorhagial nematocyst discharge and ectodermal peeling, the culmination of the response, were shown to require prior acrorhagial expansion in Anthopleura krebsi and Bunodosoma cavernata. A mechanical mechanism is suggested where- by distention of the acrorhagus opens a ciliary pit on the nematocyte surface and exposes the pit wall and microvilli, which may contain the chemoreceptors for the peeling process, including nematocyst discharge. A similar system may also be responsible for changing the threshold of nematocyst discharge in sea anemone tentacles. A case of possible neurosecretion in an anthozoan was also shown in this study.