CYSTATIN RELATED EPIDIDYMAL SPERMATOGENIC PROTEIN RESIDES IN THE OUTER DENSE FIBRES AND LIKELY TRANSIENTLY ASSOCIATES WITH THE SURFACE OF EPIDIDYMAL MOUSE SPERMATOZOA

Cystatin Related Epididymal Spermatogenic protein (CRES) is expressed in both the testis and epididymis and found associated with spermatozoa. It appears as non-glycosylated (14 and 12 kDa) and glycosylated isoforms (19 and 17 kDa). The role of CRES is enigmatic and dependent on localization of its isoforms, which is the objective of this study. The initial approach was to investigate testicular and epididymal origins of these isoforms by immunohistochemistry and immunogold cytochemistry. To further pinpoint CRES localization we then selectively extracted and fractionated epididymal spermatozoa in order to find by immunoblotting which sperm fractions contained CRES isoforms. Immunohistochemical analysis of mouse spermatogenesis showed that CRES was expressed in the tail cytoplasm of elongating spermatids from step 9-16, with a pattern reminiscent of outer dense fibre (ODF) proteins. Ultrastructural immunocytochemistry revealed that the immunogold label was concentrated over growing ODFs and mitochondrial sheath in the testes which persisted in spermatozoa through the epididymis. Sequential extractions of isolated sperm tails with Triton X-100-dithiothreitol (DTT) to remove the mitochondrial sheath, whose extract contained an unrelated 66 kDa immunoreactive band, followed by either sodium dodecyl sulfate (SDS)-DTT or urea-DTT to solubilise accessory fibres of the tail revealed a 14 kDa immunoreactive band associated with the ODF. In addition, Western blots revealed glycosylated and non-glycosylated CRES isoforms in nonyl phenoxylpolyethoxylethanol (NP40) extracts of the caput, but not cauda, sperm. Immunohistochemical analysis of the caput and cauda epithelium showed that CRES is secreted by the Golgi apparatus of the ii initial segment, fills the proximal caput lumen, and disappears by mid caput. Western blots of caput and cauda tissue and luminal fluid revealed 14 and 19 kDa immunoreactive bands in caput tissues and luminal fluid, but not in the cauda. This study concludes that there are two origins of CRES, one arising in the testis and the other in the epididymis. Testicular CRES is ionically and covalently associated with the ODF while epididymal CRES is detergent soluble and is most likely associated temporarily with the surface of caput epididymal sperm.

CERAMICS IN BRITAIN (1840-90): MEANINGS AND METAPHORS

The intention of this thesis, “Ceramics in Britain (1840–90): Meanings and Metaphors” is to present new approaches for interpreting ceramics in nineteenth-century Britain by situating, problematizing, and contextualizing pottery and porcelain in the popular debates of the day within the methodologies of material culture, design, cultural and art histories. I ask how did ceramics—portable, functional, and often decorative objects—contribute to shaping modes of experiences? Crockery, tableware and blue-white-porcelain, admittedly largely mediated in texts and paintings, are at the centre of this research to examine how they imposed symbolism and influenced the engagement of their subjects beyond their intended meanings and functions. This thesis tracks a common rhetoric shared by writers and artists across genres and understood by readers and viewers: crockery in the cupboard, on the mantel, the table or the floor were popular motifs exemplifying class, gender, character, etiquette, and taste. This thesis also seeks to map ceramics’ relations with other objects and people depicted. Their meanings and metaphors changed, depending on their exchange with other objects in the room and who uses them. The conventions of representing ceramics dictated a particular grammar that writers and artists used, critiqued, discarded or personalized. The examination of ceramics mediated in text and image especially in comparison with extant objects invites a deeper probing of both material culture and artistic practice, which helps to situate the agency of the ceramic objects themselves. Also this thesis, in attempt to explore new methodological approaches for ceramic studies, examines the social life of the mid-Victorian relief-moulded “Minster” Jug in the Gardiner Museum in Toronto. The product originating in Staffordshire in 1843 and exported to the colonies holds significance due to its multiple life histories. Viewing the “Minster” through the lenses of curator, collector, consumer, and critic its layered lives unfold to reveal the protocols of museum praxis as well as important aspects of mid-nineteenth-century British society related to design reform, gender, imperialism and consumption patterns. This thesis contends that the British experienced ceramics in sometimes unexpected ways, unrelated to their original purpose, such as tools of violence or containers of solace, and transformative fantasy.